Enhanced secretion of cyclodextrin glucanotransferase (CGTase) by Lactococcus lactis using heterologous signal peptides and optimization of cultivation conditions
| Autor: | Hafizah Mahmud, Abbas Ismail, Rosli Md. Illias, Raha Abdul Rahim, Kheng Oon Low |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Signal peptide Sequence analysis Heterologous Bioengineering Protein Sorting Signals 01 natural sciences Applied Microbiology and Biotechnology law.invention 03 medical and health sciences Bacterial Proteins law 010608 biotechnology Secretion chemistry.chemical_classification biology Lactococcus lactis General Medicine biology.organism_classification Amino acid Protein Transport 030104 developmental biology Secretory protein chemistry Biochemistry Glucosyltransferases Recombinant DNA Hydrophobic and Hydrophilic Interactions Biotechnology |
| Zdroj: | Journal of biotechnology. 296 |
| ISSN: | 1873-4863 |
| Popis: | In previous studies of Lactococcus lactis, the levels of proteins secreted using heterologous signal peptides were observed to be lower than those obtained using the signal peptide from Usp45, the major secreted lactococcal protein. In this study, G1 (the native signal peptide of CGTase) and the signal peptide M5 (mutant of the G1 signal peptide) were introduced into L. lactis to investigate the effect of signal peptides on lactococcal protein secretion to improve secretion efficiency. The effectiveness of these signal peptides were compared to the Usp45 signal peptide. The highest secretion levels were obtained using the G1 signal peptide. Sequence analysis of signal peptide amino acids revealed that a basic N-terminal signal peptide is not absolutely required for efficient protein export in L. lactis. Moreover, the introduction of a helix-breaking residue in the H-region of the M5 signal peptide caused a reduction in the signal peptide hydrophobicity and decreased protein secretion. In addition, the optimization of cultivation conditions for recombinant G1-CGTase production via response surface methodology (RSM) showed that CGTase activity increased approximately 2.92-fold from 5.01 to 16.89 U/ml compared to the unoptimized conditions. |
| Databáze: | OpenAIRE |
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